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IMPORTANCE OF HCV GENOTYPING AND
MULTIPLEX PCR FOR GENOTYPING
By, Ushanandini Mohanraj
FACTS:-
ONLY 20% OF PEOPLE INFECTED WITH HEPATITIS C VIRUS
WILL COMPLETELY RECOVER
HCV IS 10 TIMES MORE CONTAGIOUS THAN HIV WHEN
COMPARING DIRECT BLOOD-TO-BLOOD CONTACT
15 MILLION PEOPLE IN INDIA ARE AFFECTED BY HEPATITIS C
SILENT DISEASE
MORE THAN 3 LAKHS FOR CURRENT TREATMENT REGIMENT OF
24- 48 WEEKS WITH SIDE AFFECTS
WHY GENOTYPE HEPATITIS C VIRUS?
Genotype
HCV Treatment Recommendations by
AASLD
1 Harvoni for 12 weeks
2 Sovaldi + ribavirin for 12 weeks
3 Sovaldi + ribavirin for 24 weeks
4 Harvoni for 12 weeks / Sovaldi + ribavirin for
24 weeks
5 Sovaldi + ribavirin + PEG for 12 weeks
6 Harvoni for 12 weeks
Molecular methods:-
1] Innolipa
2] Restriction Fragment length
polymorphism
3] Phylogenetic analysis and sequencing
Serological method:-
1] Use of genotype specific antibodies
GENOTYPING METHODS
INNO LIPA
EXCEPT GENOTYPE 6 GOOD FOR GENOTYPING AT SUBTYPE LEVEL ONLY 1b AND 3b DISTINGUISHED ACCURATELY
PHYLOGENETIC ANALYSIS AND SEQUENCING:-
THREE STEPS:-
- IS GENOTYPING OF HEPATITIS C VIRUS BASED ON PHYLOGENETIC ANALYSIS OF 5’UTR REGION ACCURATE?
- IS THERE A NEED TO ANALYSE OTHER REGIONS OF HCV GENOME?
NS5B REGION:-
Sofosbuvir, an antiviral drug targeting NS5B, approved by the FDA in December 2013. 1.1 LAKHS FOR 24 WEEK TREATMENT Dasabuvir , Non-nucleoside NS5B polymerase inhibitor , Approved by FDA in December 2014
1000 BASES
CORE REGION:-
500 TO 600 BASES HCV core protein is a potential modulator of the host immune system
NS5B CORE
1]Specificity 2]Accurate results
1] sequencing is time consuming and a costly affair 2]sequencing of amplified DNA does not usually identify mixed infections in patients.
IDEAL GENOTYPING METHOD:-
1] ACCURATE AND RELIABLE
2] CHEAP
3] FASTER RESULTS
“GENOTYPING USING MULTIPLEX PCR”
Key features:-
1] results with just
electrophoresis
2] No need of sequencing
3] less cost and time
CHALLENGES:-
-- DECIDE WHICH REGION TO FOCUS -- TEST EFFICIENCY -- DESIGN SPECIFIC PRIMERS -- NEED FOR GENOTYPING STUDIES
IS GENOTYPING USEFUL FOR ONLY TREATMENT RECOMMEDATIONS?
-- The geographic distribution and diversity of HCV genotypes -- Tracing the source of an HCV outbreak in a given population. -- Potential Role of HCV heterogeneity and genotypes in mother-to-infant Transmission of HCV -- The role of HCV genotypes in the progression of liver disease is one of the most controversial areas of HCV research
WHATS’ MORE TO THE GENOTYPING RESEARCH?